Molecular Biotechnology最新文献

{"title":"Biogenic Silver Nanoparticles from the Cell-Free Supernatant of Mychonastes sp. B1: Antibacterial and Antibiofilm Effects, and Wound Healing Activity Supported by Gene and Protein Expression Analysis.","authors":"Enver Ersoy Andeden, Serap Nigdelioglu Dolanbay, Gulsah Avci, Belma Aslim","doi":"10.1007/s12033-026-01579-w","DOIUrl":"https://doi.org/10.1007/s12033-026-01579-w","url":null,"abstract":"<p><p>The biogenic synthesis of silver nanoparticles (AgNPs) using microalgae provides a sustainable alternative to conventional physicochemical methods. In this study, AgNPs were synthesized from the cell-free supernatant of the freshwater microalga Mychonastes sp. B1 and characterized by ultraviolet-visible spectroscopy (UV-Vis), transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), and field-emission scanning electron microscopy with energy-dispersive X-ray spectroscopy (FE-SEM/EDS). The nanoparticles were predominantly spherical (15-55 nm), highly stable (ζ =  - 42.8 mV), and appeared to be capped by extracellular polymeric substances. The biogenic AgNPs (GS-AgNPs) exhibited potent antibacterial activity, with minimum inhibitory concentrations (MICs) of 2.0 µg/mL against Staphylococcus aureus and 2.5 µg/mL against Pseudomonas aeruginosa, and significantly (p < 0.05) inhibited biofilm formation. Fibroblast viability remained at or above 80% at AgNP concentrations up to 1.5 µg/mL, which promoted cell migration and increased wound closure by 8.1% at 24 h (p < 0.05). Exposure to 1.5 µg/mL AgNPs significantly upregulated extracellular matrix markers (Col1a1 2.3-fold, Fn1 3.3-fold at mRNA level; COL1A1 2.1-fold, FN1 2.7-fold at the protein level). These findings indicate that GS-AgNPs possess antimicrobial and wound healing properties, highlighting their potential as biocompatible nanomaterials for biomedical applications.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147840564","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A CHO-Derived Matrix Attachment Region Enhances Transgene Dosage, SATB1 Recruitment, and Monoclonal Antibody Expression in a Dual-Promoter Vector System.","authors":"Sarmishta Majumdar, Rupashree Salvi, Prajakta Dandekar, Ratnesh Jain","doi":"10.1007/s12033-026-01577-y","DOIUrl":"https://doi.org/10.1007/s12033-026-01577-y","url":null,"abstract":"<p><p>The production of monoclonal antibodies (mAbs) in Chinese hamster ovary (CHO) cells is often affected by position-effect variegation and the gradual loss of transgene expression over time. Hence, we have designed a dual-promoter IgG expression vector and compared versions that either contained or lacked a CHO-derived matrix-attachment region (MAR). Stable CHO-S pools, cultured in serum-free conditions, revealed that the MAR-containing construct produced higher and more consistent antibody levels across ten passages, as confirmed by Western blot and Protein A Octet analysis. Product-quality analysis by size-exclusion chromatography and reducing SDS-PAGE confirmed formation of properly assembled, mainly monomeric antibodies in both cases. Quantitative PCR indicated greater transgene copy numbers in MAR pools (+ 48% for the light chain and + 71% for the heavy chain), and RT-qPCR showed roughly fourfold higher transcript levels for both chains relative to controls. Bioinformatic analysis revealed several SATB1 binding motifs within the MAR sequence, and ChIP-qPCR demonstrated SATB1 association with the MAR-linked transgene locus. Overall, the data suggested that a CHO-native MAR could enhance transgene dosage and transcriptional activity, while preserving product integrity, possibly through SATB1-mediated chromatin organization. Ongoing work includes chromatin-mark profiling and process-level productivity measurements to better define the impact of MAR-based vector design on biomanufacturing performance.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cloning, Transformation, and Reporter Gene Analysis of the SalT Promoter in Barley (Hordeum vulgare).","authors":"Zahid Abbas Malik, Jochen Kumlehn, Sabir Hussain Shah, Zeshan Hassan, Götz Hensel, Nasir Ahmad Saeed","doi":"10.1007/s12033-026-01570-5","DOIUrl":"https://doi.org/10.1007/s12033-026-01570-5","url":null,"abstract":"<p><p>Constitutive gene expression can lead to pleiotropic effects. Therefore, spatial or temporal restriction of expression via specific promoters provides a more targeted approach. This study aimed to clone the SalT promoter and analyze its activity in transgenic barley using GFP and GUS reporter genes. The T-DNA constructs carrying the SalT promoter were introduced into barley cv. Golden Promise, and transgenic plants were confirmed through PCR, hygromycin selection, and Southern hybridization. Both constructs, SalT-GFP and SalT-GUS, were transformed in barley cv. Golden Promise. Here, we characterized the expression pattern of the SalT promoter in barley and utilized it to drive the expression of reporter genes GFP and GUS. The SalT promoter was isolated from rice genomic DNA, cloned into the pNos-AB-M vector, and confirmed through PCR and restriction analysis. Subsequently, GFP and GUS genes were cloned under the SalT promoter in the same vector. The constructs were then subcloned into the p6U vector for plant expression. Agrobacterium-mediated genetic transformation of barley cultivar \"Golden Promise\" was conducted, resulting in successful integration of the transgenes. Callus induction, regeneration, and root formation efficiency were assessed, demonstrating the potential of the SalT promoter to drive gene expression during various stages of plant development. Molecular analyses, including PCR and Southern hybridization, confirmed the presence and integration of transgenes in the barley genome. Furthermore, GFP fluorescence and GUS staining analyses revealed strong expression of the respective genes under control of the SalT promoter in different plant tissues. This study provides insights into the application of the SalT promoter for genetic manipulation and functional characterization in barley, offering opportunities for crop improvement and biotechnological applications.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MicroRNA Crosstalk in Metabolic Disorders: The Dual Role of miR-122 and miR-34a in NAFLD and Type 2 Diabetes.","authors":"Sumita Thakur, Anjana Munshi, Prabhsimran Kaur","doi":"10.1007/s12033-026-01576-z","DOIUrl":"https://doi.org/10.1007/s12033-026-01576-z","url":null,"abstract":"<p><p>NAFLD and T2DM are metabolic diseases with overlapping pathogenic mechanisms, including insulin resistance, lipid imbalance, and chronic inflammation. miRNAs have emerged as reliable biomarkers for NAFLD and T2DM. The expression levels of miRNAs hold the potential to contribute to bridging the gap between metabolic diseases. miR-122 and miR-34a are significant regulators of metabolic pathways among various biomarkers. miR-122, a liver-enriched microRNA involved in hepatic lipid metabolism, inflammation, and insulin signaling, whereas miR-34a is closely linked to β-cell dysfunction and hepatic steatosis through SIRT1 suppression. Growing echelons of these miRNAs in the bloodstream of patients with T2DM and NAFLD show their potential use as prognostic and diagnostic indicators. These enable a non-invasive early disease monitoring and detection alternative. miR-122 and miR-34a are involved in disease progression by influencing key metabolic processes such as lipid metabolism, pro-inflammatory cytokine regulation, the SIRT1-SREBP1c pathway, and AMPK phosphorylation. Their changed expression levels in various metabolic conditions contribute to advancing pathophysiological processes, explaining their clinical value as therapeutic targets. This review investigates the molecular functions of miR-122 and miR-34a in NAFLD and T2DM, emphasizing their effect on various genes and interrelated pathways, showing their diagnostic potential and therapeutic implications. Knowing their regulatory roles might lead the way for miRNA-based precision medicine techniques that improve early diagnosis, risk assessment, and treatment strategies for metabolic illnesses. Future research should focus on their therapeutic use and make targeted interventions to change their expression levels for disease management.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147776846","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research Advances in Multi-tissue Organoid Models Based on PBMCs.","authors":"Dongyang He, Yicheng Feng, Xiao An","doi":"10.1007/s12033-026-01573-2","DOIUrl":"https://doi.org/10.1007/s12033-026-01573-2","url":null,"abstract":"<p><p>Organoids, as three-dimensional in vitro culture models, closely recapitulate the architectural, cellular, and functional characteristics of their in vivo counterparts. Consequently, they play a pivotal role in diverse biomedical fields, including disease modeling, drug screening, and regenerative medicine. However, conventional organoid models often lack a complex native microenvironment, specifically the tumor microenvironment (TME), which limits their utility in studying immune-related pathologies and evaluating immunotherapeutic strategies. Peripheral blood mononuclear cells (PBMCs) offer distinct advantages, including accessibility, a heterogeneous composition of immune cells, and the capacity to mirror a patient's specific immune status. As a result, PBMCs have emerged as a predominant source of immune cells for engineering immune-integrated organoid models. This article reviews recent advancements in integrating PBMCs into various tissue organoid systems. It summarizes the key biological properties of PBMCs and their applications across multiple models, including cerebral, renal, pulmonary, and hepatic organoids. Furthermore, this study compares the advantages and limitations of current modeling methodologies and discusses future research directions and persistent challenges in leveraging PBMC-integrated organoids for personalized medicine and the elucidation of disease mechanisms.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147776832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of an Efficient Regeneration and Agrobacterium-Mediated Transformation Protocol for Hosta 'Light Star' Using the RUBY Reporter Gene.","authors":"Hangkuan Zhou, Yidi Qin, Shuang Liu, Xinyue Zhang, Chen Chen, Jinzhu Zhang, Tao Yang, Wuhua Zhang, Jinping Fan","doi":"10.1007/s12033-026-01575-0","DOIUrl":"https://doi.org/10.1007/s12033-026-01575-0","url":null,"abstract":"<p><p>Hosta plantaginea is a perennial shade-tolerant herb of the Liliaceae family, with high ornamental and urban greening value. Hosta 'Light Star' is a newly developed ornamental cultivar with yellow-margined leaves and lilac flowers, but no efficient in vitro regeneration or genetic transformation system has been established for this cultivar to date. In this study, we established a highly efficient in vitro regeneration system for Hosta 'Light Star,' and developed an Agrobacterium-mediated genetic transformation protocol using the RUBY visual reporter gene for non-invasive screening of positive transformants. The optimal callus induction medium was MS + 2 mg/L 6-BA + 0.3 mg/L NAA + 0.05 mg/L 2, 4-D, with a callus induction rate of 53.33% for leaf explants (the optimal explant for sterile seedlings). The optimal adventitious bud proliferation medium was MS + 2 mg/L 6-BA + 0.1 mg/L NAA, with a proliferation coefficient of 5.87. The optimal rooting medium was 1/2 MS + 0.5 mg/L NAA + 0.5 mg/L IBA, with a 100% rooting rate. The optimal transplant substrate was perlite:vermiculite = 2:1, with a 100% transplant survival rate after acclimatization. For Agrobacterium-mediated transformation, the optimal infection parameters were as follows: Agrobacterium suspension OD₆₀₀ = 0.6, infection time of 10 min, and 200 μM acetosyringone; the optimal selection conditions were 300 mg/L cefotaxime for bacteriostasis and 30 mg/L hygromycin for transformant screening. The final stable transformation efficiency was 2.50% (95% CI 1.23-3.77%), with an escape rate of 16.13%. Transgenic plants showed distinct purplish-red coloration in roots, stems, and leaves, with significantly higher betacyanin accumulation than wild-type plants (p < 0.05). Stable integration and expression of the RUBY gene were confirmed by PCR, RT-PCR, and RT-qPCR. This study establishes the first efficient regeneration and Agrobacterium-mediated transformation system for Hosta 'Light Star,' and validates the feasibility of the RUBY reporter gene as a visual marker for Hosta transformation. This system provides a solid technical platform for functional genomic studies, CRISPR/Cas9-mediated gene editing, and molecular breeding of ornamental traits in Hosta.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147729513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Comprehensive Review on CRISPR-Based Screening and Its Applications.","authors":"Ali Saber Sichani, Mahmoud Hassani, Fatemeh Gila, Negin Shafieipour, Reza Dabbaghipour, Zahra Heidari, Mohsen Sisakht, Mohammad Hassani, Jafar Fallahi","doi":"10.1007/s12033-026-01562-5","DOIUrl":"https://doi.org/10.1007/s12033-026-01562-5","url":null,"abstract":"<p><p>CRISPR-based tools have quickly moved from specialist techniques to routine instruments in biology and medicine, and they are now central to large-scale loss-of-function and perturbation screens. In this review, we focus on how pooled CRISPR screens are used to interrogate gene function in living cells, most often through cell fitness or simple selectable markers, and contrast this with arrayed formats that trade throughput for richer molecular readouts, such as transcriptome-wide changes. We bring together current strategies for library design, delivery, and selection and show how different Cas nucleases, including Cas9, Cas12, and Cas13, broaden the range of genome and transcriptome perturbations that can be assayed. We then discuss recent applications in drug response, viral infection, and cancer biology and consider how improvements in high-content technologies, data analysis, and emerging diagnostic uses are likely to shape the next generation of CRISPR-based screening studies.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147723370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microbiological Profile and Antimicrobial Resistance Patterns of Pleural Fluid Isolates in a Tertiary Care Setting in Lahore, Pakistan.","authors":"Hassan Imran, Zaman Khan, Fatima Iftikhar Shah, Fatima Akram, Fiza Saleem, Somia Shehzadi","doi":"10.1007/s12033-026-01574-1","DOIUrl":"https://doi.org/10.1007/s12033-026-01574-1","url":null,"abstract":"<p><p>Pleural fluid infections, primarily caused by a range of bacterial pathogens, present a growing challenge for healthcare professionals, particularly due to the increasing prevalence of antibiotic resistance. This study included 598 patients, of which 15.88% had culture-positive samples. A total of 18 distinct bacterial species, both individual and in combination, were identified, with 7 Gram-positive and 11 Gram-negative bacteria. Antibacterial susceptibility testing highlighted colistin as the most effective antibiotic for most Gram-negative pathogens, while vancomycin and linezolid showed increased sensitivity against Gram-positive isolates. The study calculated the Multiple Antibiotic Resistance (MAR) index for all isolates, yielding a mean value of 0.543, reflecting the high level of irrational antibiotic use. Resistance pattern analysis revealed significant variation among bacterial species and identified 61 distinct resistance profiles. The study also examined the relationship between the MAR index and factors such as gender, age, and Gram staining. Gender (p = 0.831) and age (p = 0.905) showed no significant correlation with the MAR index, while a significant negative correlation was found between Gram staining and the MAR index (ρ = - 0.434, p < 0.001). The findings highlight the increasing prevalence of multidrug-resistant (MDR) and extensively drug-resistant (XDR) isolates, presenting significant challenges for treatment, particularly in resource-limited settings. This underscores the urgent need for enhanced antibiotic stewardship, public awareness campaigns, and continuous monitoring to combat antibiotic resistance and improve patient outcomes. Furthermore, the study's findings contribute to global efforts in understanding and addressing the escalating problem of antibiotic resistance, offering insights relevant to international research on antimicrobial resistance.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":""},"PeriodicalIF":2.5,"publicationDate":"2026-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147618785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanoemulsion-Based Tea Tree Oil: A New Frontier in Enhancing Clotrimazole Antifungal Activity.","authors":"Fatemeh Khorsand, Seyed Jamal Hashemi, Shahram Mahmoudi, Hasti Kamali Sarvestani, Amir Amani","doi":"10.1007/s12033-025-01485-7","DOIUrl":"10.1007/s12033-025-01485-7","url":null,"abstract":"<p><p>Clotrimazole is widely used to treat fungal diseases, but it should be applied topically for several weeks to be effective. Previous studies have shown that nanoemulsions can enhance the topical activity of various active ingredients. This study aimed to prepare a nanoformulation of clotrimazole to improve its antifungal activity. A nanoemulsion containing clotrimazole was prepared using Tween 80 and Tween 20. The particle size was measured using Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM). Stability studies were conducted using freeze-thaw cycles and storage at 40 °C. The antifungal activity of clotrimazole was then compared with bulk clotrimazole both in vitro and in vivo. The nanoemulsion containing clotrimazole had a particle size of 31.5 nm, measured by DLS and remained physically stable after 1 year of storage. In vitro antifungal studies showed that this nanoemulsion outperformed both bulk clotrimazole and the nanoemulsion without clotrimazole, with MIC values of 1/524288, 1/8192, and 1/1024, respectively, against T. mentagrophytes. In addition, animal studies demonstrated that mean (SD) treatment time for fungal infection was 8.6 (0.5) days with the nanoemulsion containing 1% clotrimazole, significantly smaller than that of bulk clotrimazole (1% w/w) and nanoemulsion without clotrimazole (i.e., 15.0 (1.6) and 20.0 (0.7) days, respectively). The treatment time was more than 21 days for the bulk tea tree oil and the untreated control. After 21 days of treatment, only animals treated with the clotrimazole nanoemulsion showed complete lesion healing and weight changes comparable to healthy animals. The nanoemulsion formulation containing clotrimazole demonstrates highly desirable antifungal properties, warranting further investigation in clinical trials.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"1921-1928"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144732383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of a Phage Display-Selected scFv Against Neuropilin 1 (NRP1) Isolated from a Naïve Human Library.","authors":"Khadijeh Farrokhi, Fatemeh Nasiri, Pouya Safarzadeh Kozani, Pooria Safarzadeh Kozani","doi":"10.1007/s12033-025-01480-y","DOIUrl":"10.1007/s12033-025-01480-y","url":null,"abstract":"<p><p>Immune checkpoint inhibitors are established therapeutics for solid tumors. This study isolated a single-chain fragment variable (scFv) targeting neuropilin 1 (NRP1), a promising therapeutic target. A naïve human scFv phage display library underwent five biopanning rounds against NRP1. The lead clone (KZC28) was expressed and characterized. Binding affinity was quantified via ELISA. Specificity was assessed by cross-reactivity testing against structurally similar/unrelated antigens. Sensitivity determined the lower detection limit. Cell-based ELISA and flow cytometry evaluated binding to endogenous NRP1 on U87-MG glioblastoma and PC3 prostate cancer cells. Computational modeling predicted structural stability, solubility, flexibility, and binding energetics, focusing on complementarity-determining region (CDR) interactions. KZC28 exhibited high affinity (K_d = 0.88 nM) and specificity, with no cross-reactivity despite high sequence identity. Sensitivity reached 20 ng/mL for NRP1 detection. Cell-based assays and flow cytometry confirmed robust binding to U87-MG and PC3 cells, with specific binding comparable to a commercial anti-NRP1 mAb and minimal binding to NRP1⁻ cells. In silico analyses predicted favorable solubility, structural flexibility, and energetically stable CDR-mediated binding. KZC28 demonstrates high specificity and affinity for NRP1 with promising biophysical and functional properties. These support its potential as a targeting moiety for solid tumor therapy, particularly in NRP1-overexpressing cancers. Further preclinical evaluation of in vivo efficacy is warranted.</p>","PeriodicalId":18865,"journal":{"name":"Molecular Biotechnology","volume":" ","pages":"1881-1901"},"PeriodicalIF":2.5,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144708142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}